PXD046160 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Residues 1 to 563 of α-hemolysin alone form a fully functional pore and their proper membrane insertion depends on the acylation status of the molecule |
Description | The acylated pore-forming Repeats in ToXin (RTX) cytolysins α-hemolysin (HlyA) and adenylate cyclase toxin (CyaA) bind primarily to β2 integrins of leukocytes. HlyA binds the common CD18 subunit of the β2 integrins and CyaA selectively binds the CD11b subunit of complement receptor 3 (CR3). However, both toxins can also bind and permeabilize membranes of a variety of nonmyeloid cells. We constructed HlyA1-563/CyaA860-1706 hybrid molecules activated by the CyaA-activating acyltransferase CyaC, or by the HlyA-activating acyltransferase HlyC. We show that the C-terminal portion of the HlyA molecule, comprising the acylated segment and the RTX domain (residues 564 to 1024), can be functionally swapped with the CyaC-activated acylated segment bearing palmitoylated Lys983 and the much larger RTX domain of CyaA. Compared to the CD18-interacting HlyA, the CR3-interacting HlyA1-563/CyaA860-1706 hybrid exhibited a selectively reduced cytotoxicity on human THP-1 monocytes and Chinese hamster ovary (CHO) cells expressing CR3 (CHO-CR3) or lymphocyte function-associated antigen 1 (CHO-LFA1). However, the mono-palmitoylated HlyA1-563/CyaA860-1706 hybrid remained fully hemolytic and cytolytic toward erythrocytes and mock-transfected CHO cells and exhibited a comparable membrane activity on artificial planar lipid membranes as the bi-myristoylated HlyA. Thus, regardless of the length or number of the attached fatty acyl chains, the CyaC-activated HlyA1-563/CyaA860-1706 hybrid and the intact HlyA were comparably active on cells or artificial membranes lacking β2 integrins. These data suggest that once the hydrophobic pore-forming domain comprising N-terminal half of HlyA is brought into close contact with the cell membrane by the mono-palmitoylated acylated segment of CyaA, it can efficiently form fully cytolytic HlyA-like membrane pores. |
HostingRepository | PRIDE |
AnnounceDate | 2024-03-25 |
AnnouncementXML | Submission_2024-03-25_08:12:33.855.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | David Jurnečka |
SpeciesList | scientific name: Escherichia coli; NCBI TaxID: 562; |
ModificationList | palmitoylated residue; myristoylated residue |
Instrument | Bruker Daltonics solarix series |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2023-10-16 08:05:02 | ID requested | |
⏵ 1 | 2024-03-25 08:12:34 | announced | |
Publication List
Dataset with its publication pending |
Keyword List
submitter keyword: acylation,RTX toxin |
Contact List
Laboratory of Molecular Biology of Bacterial Pathogens |
contact affiliation | Institute of Microbiology of the Czech Academy of Sciences, Prague, Czech Republic |
contact email | sebo@biomed.cas.cz |
lab head | |
David Jurnečka |
contact affiliation | Institute of Microbiology, Czech Academy of Sciences |
contact email | david.jurnecka@biomed.cas.cz |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD046160
- Label: PRIDE project
- Name: Residues 1 to 563 of α-hemolysin alone form a fully functional pore and their proper membrane insertion depends on the acylation status of the molecule